Raw sequence reads of ADSCs in Rattus norvegicus (rat)

The aim of this study was to find a core of the proliferative subpopulation with the most stemness characteristics. Firstly, the transcriptomes of 15,532 single ADSCs were profiled using scRNA-seq. Based on gene expression patterns, 15 cell subpopulations (6 defined cell types) were distinguished, in which 37.68% were stem and progenitor cells. One of these subpopulations, subpopulation 11 was identified as CD168+ADSCs. In the subsequent studies, CD168+ADSCs were demonstrated to be in the prophase of cell mitosis. And results of qRT-PCR showed that CD168+ADSCs expressed higher levels of G2/M featured genes. Furthermore, Hmmr, a specific marker gene of CD168+ADSCs, was overexpressed and knocked out respectively by shRNA lentivirus and CRISPR/Cas9 KO plasmids. Hmmr was proved to promote ADSCs proliferation, stemness maintenance and regulate cell cycle conversion. Finally, the results of western blot and co-immunoprecipitation suggested that Hmmr promoted ADSCs proliferation through extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. Our study identified a vital role of CD168+ subpopulation in ADSCs proliferation and stemness maintenance, and suggested that CD168+ADSCs may be a useful target in stem cells and regenerative medicine.